Purpose: To introduce punch biopsy, which can be easily performed in an outpatient setting, and to evaluate its clinical usefulness. Methods: A retrospective study was performed in 33 patients with atypical eyelid disorders who underwent punch biopsy. Results: This study included 33 eyes (33 patients; male:female = 12:21) with a mean age of 57.27 ± 19.12 years. There were 19 (57.6%) right eye, 14 (42.4%) left eye, 15 (45.5%) upper eyelid, and 18 (54.5%) lower eyelid lesions. The biopsy approaches were percutaneous and transconjunctival in 24 (72.7%) and nine (27.3%) eyes, respectively. Benign and malignant lesions were found in 21 (63.6%) and 12 (36.4%) eyes, respectively. The benign lesions included chalazion (52.4%), acute and chronic inflammation (23.8%), acanthosis (4.8%), and ectopic cillia (4.8%), and were treated conservatively by intralesional steroid injections or incision and drainage. The malignant lesions included sebaceous carcinomas (33.3%), squamous cell carcinomas (25%), and basal cell carcinomas (16.7%), with various treatments, including surgical resections, chemotherapy, and radiation therapy. Conclusions Punch biopsy is a safe and quick procedure that can easily be performed in the office setting for patients who require a pathologic diagnosis for eyelid diseases, including malignancies. It may be useful for diagnosis and treatment planning.